Ants can influence ecological processes, such as the transfer of elements or radionuclides, in several ways. For example, they redistribute materials while foraging and maintaining their nests and have an important role in terrestrial food webs. Quantitative data of the transfer of elements into ants is needed, e.g., for developing improved radioecological models. In this study, samples of red wood ants (genus Formica), nest material, litter and soil were collected from a former uranium mining site in Eastern Finland. Concentrations of 33 elements were analyzed by Inductively Coupled Plasma-Mass Spectroscopy/Optical Emission Spectroscopy. Estimated element concentrations in spruce needles were used as a proxy for studying the transfer of elements into ants via aphids because spruces host the most important aphid farms in boreal forests. Empirically determined organism/medium concentration ratios (CRs) are commonly used in radioecological models. Ant/soil CRs were calculated and the validity of the fundamental assumption behind the of use of CRs (linear transfer) was evaluated. Elements that accumulated in ants in comparison to other compartments were cadmium, potassium, phosphorus, sulfur, and zinc. Ant uranium concentrations were low in comparison to soil, litter, or nest material but slightly elevated in comparison to spruce needles. Ant element concentrations were quite constant regardless of the soil concentrations. Non-linear transfer models could therefore describe the soil-to-ant transfer better than conventional CRs.

Ants can influence ecological processes, such as the transfer of elements or radionuclides, in several ways. For example, they redistribute materials while foraging and maintaining their nests and have an important role in terrestrial food webs. Quantitative data of the transfer of elements into ants is needed, e.g., for developing improved radioecological models. In this study, samples of red wood ants (genus Formica), nest material, litter and soil were collected from a former uranium mining site in Eastern Finland. Concentrations of 33 elements were analyzed by Inductively Coupled Plasma-Mass Spectroscopy/Optical Emission Spectroscopy. Estimated element concentrations in spruce needles were used as a proxy for studying the transfer of elements into ants via aphids because spruces host the most important aphid farms in boreal forests. Empirically determined organism/medium concentration ratios (CRs) are commonly used in radioecological models. Ant/soil CRs were calculated and the validity of the fundamental assumption behind the of use of CRs (linear transfer) was evaluated. Elements that accumulated in ants in comparison to other compartments were cadmium, potassium, phosphorus, sulfur, and zinc. Ant uranium concentrations were low in comparison to soil, litter, or nest material but slightly elevated in comparison to spruce needles. Ant element concentrations were quite constant regardless of the soil concentrations. Non-linear transfer models could therefore describe the soil-to-ant transfer better than conventional CRs. | 2022

The presence of contaminants of emerging concern in the aquatic environment directly impacts water-living organisms and can alter their living functions. These compounds are often metabolized and excreted, but they can also be accumulated and spread through the food chain. The metabolized contaminants can also lead to the formation of new compounds with unknown toxicity and bioaccumulation potential. In this work, we have studied the occurrence, bioconcentration, and biotransformation of CECs in glass eels (Anguilla anguilla) using UHPLC-HRMS. To select the target CECs, we first carried out an environmental risk assessment of the WWTP effluent that releases directly into the Adour estuary (Bayonne, Basque Country, France). The risk quotients of every detected contaminant were calculated and three ecotoxicologically relevant contaminants were chosen to perform the exposure experiment: propranolol, diazepam, and irbesartan. An experiment of 14 days consisting of 7 days of exposure and 7 days of depuration was carried out to measure the bioconcentration of the chosen compounds. The quantitative results of the concentrations in glass eel showed that diazepam and irbesartan reached BCF ≈10 on day 7, but both compounds were eliminated after 7 days of depuration. On the other hand, propranolol's concentration remains constant all along with the experiment, and its presence can be detected even in the non-exposed control group, which might suggest environmental contamination. Two additional suspect screening strategies were used to identify metabolization products of the target compounds and other xenobiotics already present in wild glass eels. Only one metabolite was identified, nordiazepam, a well-known diazepam metabolite, probably due to the low metabolic rate of glass eels at this stage. The xenobiotic screening confirmed the presence of more xenobiotics in wild glass eels, prominent among them, the pharmaceuticals exemestane, primidone, iloprost, and norethandrolone.

International audience | The presence of contaminants of emerging concern in the aquatic environment directly impacts water-living organisms and can alter their living functions. These compounds are often metabolized and excreted, but they can also be accumulated and spread through the food chain. The metabolized contaminants can also lead to the formation of new compounds with unknown toxicity and bioaccumulation potential. In this work, we have studied the occurrence, bioconcentration, and biotransformation of CECs in glass eels (Anguilla anguilla) using UHPLC-HRMS. To select the target CECs, we first carried out an environmental risk assessment of the WWTP effluent that releases directly into the Adour estuary (Bayonne, Basque Country, France). The risk quotients of every detected contaminant were calculated and three ecotoxicologically relevant contaminants were chosen to perform the exposure experiment: propranolol, diazepam, and irbesartan. An experiment of 14 days consisting of 7 days of exposure and 7 days of depuration was carried out to measure the bioconcentration of the chosen compounds. The quantitative results of the concentrations in glass eel showed that diazepam and irbesartan reached BCF ≈10 on day 7, but both compounds were eliminated after 7 days of depuration. On the other hand, propranolol's concentration remains constant all along with the experiment, and its presence can be detected even in the non-exposed control group, which might suggest environmental contamination. Two additional suspect screening strategies were used to identify metabolization products of the target compounds and other xenobiotics already present in wild glass eels. Only one metabolite was identified, nordiazepam, a well-known diazepam metabolite, probably due to the low metabolic rate of glass eels at this stage. The xenobiotic screening confirmed the presence of more xenobiotics in wild glass eels, prominent among them, the pharmaceuticals exemestane, primidone, iloprost, and norethandrolone. ☆ This paper has been recommended for acceptance by. Eddy Y. Zeng. ☆☆ Contaminants of Emerging Concern in Glass Eel (Anguilla anguilla): Occurrence, Bioconcentration and Biotransformation.

Nanosilver (NAg) is currently one of the major alternative antimicrobials to control microorganisms. With its broad-spectrum efficacy and lucrative commercial values, NAg has been used in medical devices and increasingly, in consumer products and appliances. This widespread use has inevitably led to the release and accumulation of the nanoparticle in water and sediment, in soil and even, wastewater treatment plants (WWTPs). This Article describes the physical and chemical transformations of NAg as well as the impact of the nanoparticle on microbial communities in different environmental settings; how the nanoparticle shifts not only the diversity and abundance of microbes, including those that are important in nitrogen cycles and decomposition of organic matters, but also their associated genes and in turn, the key metabolic processes. Current findings on the microbiological activity of the leached soluble silver, solid silver particulates and their respective transformed products, which underpin the mechanism of the nanoparticle toxicity in environmental microbes, is critically discussed. The Article also addresses the emerging evidence of silver-driven co-selection of antibiotic resistance determinants. The mechanism has been linked to the increasing pools of many antibiotic resistance genes already detected in samples from different environmental settings, which could ultimately find their ways to animals and human. The realized ecological impact of NAg calls for more judicial use of the nanoparticle. The generated knowledge can inform strategies for a better ‘risks versus benefits’ assessment of NAg applications, including the disposal stage.

Nanosilver (NAg) is currently one of the major alternative antimicrobials to control microorganisms. With its broad-spectrum efficacy and lucrative commercial values, NAg has been used in medical devices and increasingly, in consumer products and appliances. This widespread use has inevitably led to the release and accumulation of the nanoparticle in water and sediment, in soil and even, wastewater treatment plants (WWTPs). This Article describes the physical and chemical transformations of NAg as well as the impact of the nanoparticle on microbial communities in different environmental settings; how the nanoparticle shifts not only the diversity and abundance of microbes, including those that are important in nitrogen cycles and decomposition of organic matters, but also their associated genes and in turn, the key metabolic processes. Current findings on the microbiological activity of the leached soluble silver, solid silver particulates and their respective transformed products, which underpin the mechanism of the nanoparticle toxicity in environmental microbes, is critically discussed. The Article also addresses the emerging evidence of silver-driven co-selection of antibiotic resistance determinants. The mechanism has been linked to the increasing pools of many antibiotic resistance genes already detected in samples from different environmental settings, which could ultimately find their ways to animals and human. The realized ecological impact of NAg calls for more judicial use of the nanoparticle. The generated knowledge can inform strategies for a better 'risks versus benefits' assessment of NAg applications, including the disposal stage. | Kevin Yonathan, Riti Mann, Khandaker Rayhan Mahbub, Cindy Gunawan

Nanosilver (NAg) is currently one of the major alternative antimicrobials to control microorganisms. With its broad-spectrum efficacy and lucrative commercial values, NAg has been used in medical devices and increasingly, in consumer products and appliances. This widespread use has inevitably led to the release and accumulation of the nanoparticle in water and sediment, in soil and even, wastewater treatment plants (WWTPs). This Article describes the physical and chemical transformations of NAg as well as the impact of the nanoparticle on microbial communities in different environmental settings; how the nanoparticle shifts not only the diversity and abundance of microbes, including those that are important in nitrogen cycles and decomposition of organic matters, but also their associated genes and in turn, the key metabolic processes. Current findings on the microbiological activity of the leached soluble silver, solid silver particulates and their respective transformed products, which underpin the mechanism of the nanoparticle toxicity in environmental microbes, is critically discussed. The Article also addresses the emerging evidence of silver-driven co-selection of antibiotic resistance determinants. The mechanism has been linked to the increasing pools of many antibiotic resistance genes already detected in samples from different environmental settings, which could ultimately find their ways to animals and human. The realized ecological impact of NAg calls for more judicial use of the nanoparticle. The generated knowledge can inform strategies for a better 'risks versus benefits' assessment of NAg applications, including the disposal stage.

Increasing terrestrial run-off from melting glaciers and thawing permafrost to Arctic coastal areas is expected to facilitate re-mobilization of stored legacy persistent organic pollutants (POPs) and mercury (Hg), potentially increasing exposure to these contaminants for coastal benthic organisms. We quantified chlorinated POPs and Hg concentrations, lipid content and multiple dietary markers, in a littoral deposit-feeding amphipod Gammarus setosus and sediments during the melting period from April to August in Adventelva river estuary in Svalbard, a Norwegian Arctic Aarchipelago. There was an overall decrease in concentrations of ∑POPs from April to August (from 58 ± 23 to 13 ± 4 ng/g lipid weight; lw), Hg (from 5.6 ± 0.7 to 4.1 ± 0.5 ng/g dry weight; dw) and Methyl Hg (MeHg) (from 5 ± 1 to 0.8 ± 0.7 ng/g dw) in G. setosus. However, we observed a seasonal peak in penta- and hexachlorobenzene (PeCB and HCB) in May (2.44 ± 0.3 and 23.6 ± 1.7 ng/g lw). Sediment concentrations of POPs and Hg (dw) only partly correlated with the contaminant concentrations in G. setosus. Dietary markers, including fatty acids and carbon and nitrogen stable isotopes, indicated a diet of settled phytoplankton in May–July and a broader range of carbon sources after the spring bloom. Phytoplankton utilization and chlorobenzene concentrations in G. setosus exhibited similar seasonal patterns, suggesting a dietary uptake of chlorobenzenes that is delivered to the aquatic environment during spring snowmelt. The seasonal decrease in contaminant concentrations in G. setosus could be related to seasonal changes in dietary contaminant exposure and amphipod ecology. Furthermore, this decrease implies that terrestrial run-off is not a significant source of re-mobilized Hg and legacy POPs to littoral amphipods in the Adventelva river estuary during the melt season.

Increasing terrestrial run-off from melting glaciers and thawing permafrost to Arctic coastal areas is expected to facilitate re-mobilization of stored legacy persistent organic pollutants (POPs) and mercury (Hg), potentially increasing exposure to these contaminants for coastal benthic organisms. We quantified chlorinated POPs and Hg concentrations, lipid content and multiple dietary markers, in a littoral deposit-feeding amphipod Gammarus setosus and sediments during the melting period from April to August in Adventelva river estuary in Svalbard, a Norwegian Arctic Aarchipelago. There was an overall decrease in concentrations of ∑POPs from April to August (from 58 ± 23 to 13 ± 4 ng/g lipid weight; lw), Hg (from 5.6 ± 0.7 to 4.1 ± 0.5 ng/g dry weight; dw) and Methyl Hg (MeHg) (from 5 ± 1 to 0.8 ± 0.7 ng/g dw) in G. setosus. However, we observed a seasonal peak in penta- and hexachlorobenzene (PeCB and HCB) in May (2.44 ± 0.3 and 23.6 ± 1.7 ng/g lw). Sediment concentrations of POPs and Hg (dw) only partly correlated with the contaminant concentrations in G. setosus. Dietary markers, including fatty acids and carbon and nitrogen stable isotopes, indicated a diet of settled phytoplankton in May–July and a broader range of carbon sources after the spring bloom. Phytoplankton utilization and chlorobenzene concentrations in G. setosus exhibited similar seasonal patterns, suggesting a dietary uptake of chlorobenzenes that is delivered to the aquatic environment during spring snowmelt. The seasonal decrease in contaminant concentrations in G. setosus could be related to seasonal changes in dietary contaminant exposure and amphipod ecology. Furthermore, this decrease implies that terrestrial run-off is not a significant source of re-mobilized Hg and legacy POPs to littoral amphipods in the Adventelva river estuary during the melt season. | publishedVersion

Increasing terrestrial run-off from melting glaciers and thawing permafrost to Arctic coastal areas is expected to facilitate re-mobilization of stored legacy persistent organic pollutants (POPs) and mercury (Hg), potentially increasing exposure to these contaminants for coastal benthic organisms. We quantified chlorinated POPs and Hg concentrations, lipid content and multiple dietary markers, in a littoral deposit-feeding amphipod Gammarus setosus and sediments during the melting period from April to August in Adventelva river estuary in Svalbard, a Norwegian Arctic Aarchipelago. There was an overall decrease in concentrations of ∑POPs from April to August (from 58 ± 23 to 13 ± 4 ng/g lipid weight; lw), Hg (from 5.6 ± 0.7 to 4.1 ± 0.5 ng/g dry weight; dw) and Methyl Hg (MeHg) (from 5 ± 1 to 0.8 ± 0.7 ng/g dw) in G. setosus. However, we observed a seasonal peak in penta- and hexachlorobenzene (PeCB and HCB) in May (2.44 ± 0.3 and 23.6 ± 1.7 ng/g lw). Sediment concentrations of POPs and Hg (dw) only partly correlated with the contaminant concentrations in G. setosus. Dietary markers, including fatty acids and carbon and nitrogen stable isotopes, indicated a diet of settled phytoplankton in May–July and a broader range of carbon sources after the spring bloom. Phytoplankton utilization and chlorobenzene concentrations in G. setosus exhibited similar seasonal patterns, suggesting a dietary uptake of chlorobenzenes that is delivered to the aquatic environment during spring snowmelt. The seasonal decrease in contaminant concentrations in G. setosus could be related to seasonal changes in dietary contaminant exposure and amphipod ecology. Furthermore, this decrease implies that terrestrial run-off is not a significant source of re-mobilized Hg and legacy POPs to littoral amphipods in the Adventelva river estuary during the melt season.

Increasing terrestrial run-off from melting glaciers and thawing permafrost to Arctic coastal areas is expected to facilitate re-mobilization of stored legacy persistent organic pollutants (POPs) and mercury (Hg), potentially increasing exposure to these contaminants for coastal benthic organisms. We quantified chlorinated POPs and Hg concentrations, lipid content and multiple dietary markers, in a littoral deposit-feeding amphipod Gammarus setosus and sediments during the melting period from April to August in Adventelva river estuary in Svalbard, a Norwegian Arctic Aarchipelago. There was an overall decrease in concentrations of ∑POPs from April to August (from 58 ± 23 to 13 ± 4 ng/g lipid weight; lw), Hg (from 5.6 ± 0.7 to 4.1 ± 0.5 ng/g dry weight; dw) and Methyl Hg (MeHg) (from 5 ± 1 to 0.8 ± 0.7 ng/g dw) in G. setosus. However, we observed a seasonal peak in penta- and hexachlorobenzene (PeCB and HCB) in May (2.44 ± 0.3 and 23.6 ± 1.7 ng/g lw). Sediment concentrations of POPs and Hg (dw) only partly correlated with the contaminant concentrations in G. setosus. Dietary markers, including fatty acids and carbon and nitrogen stable isotopes, indicated a diet of settled phytoplankton in May–July and a broader range of carbon sources after the spring bloom. Phytoplankton utilization and chlorobenzene concentrations in G. setosus exhibited similar seasonal patterns, suggesting a dietary uptake of chlorobenzenes that is delivered to the aquatic environment during spring snowmelt. The seasonal decrease in contaminant concentrations in G. setosus could be related to seasonal changes in dietary contaminant exposure and amphipod ecology. Furthermore, this decrease implies that terrestrial run-off is not a significant source of re-mobilized Hg and legacy POPs to littoral amphipods in the Adventelva river estuary during the melt season. | publishedVersion

Indoor dust has been postulated as an important matrix for residential pesticide exposure. However, there is a lack of information on presence, concentrations and determinants of multiple pesticides in dust in residential homes close to treated fields. Our objective was to characterize the spatial and temporal variance of pesticides in house dust, study the use of doormats and floors as proxies for pesticides in indoor dust and identify determinants of occurrence and concentrations. Homes within 250 m from selected bulb fields were invited to participate. Homes within 20 km from these fields but not having agricultural fields within 500 m were selected as controls. House dust was vacuumed in all homes from floors (VFD) and from newly placed clean doormats (DDM). Sampling was done during two periods, when pesticides are used and not-used. For determination of 46 prioritized pesticides, a multi-residue extraction method was used. Most statistical analyses are focused on the 12 and 14 pesticides that were detected in >40% of DDM and VFD samples, respectively. Mixed models were used to evaluate relationships between possible determinants and pesticides occurrence and concentrations in DDM and VFD. 17 pesticides were detected in more than 50% of the homes in both matrixes. Concentrations differed by about a factor five between use and non-use periods among homes within 250 m of fields and between these homes and controls. For 7 pesticides there was a moderate to strong correlation (Spearman rho 0.30–0.75) between concentrations in DDM and VFD. Distance to agricultural fields and air concentrations were among the most relevant predictors for occurrence and levels of a given pesticide in DDM. Concentrations in dust are overall higher during application periods and closer to fields (<250 m) than further away. The omnipresence of pesticides in dust lead to residents being exposed all year round.

Indoor dust has been postulated as an important matrix for residential pesticide exposure. However, there is a lack of information on presence, concentrations and determinants of multiple pesticides in dust in residential homes close to treated fields. Our objective was to characterize the spatial and temporal variance of pesticides in house dust, study the use of doormats and floors as proxies for pesticides in indoor dust and identify determinants of occurrence and concentrations. Homes within 250 m from selected bulb fields were invited to participate. Homes within 20 km from these fields but not having agricultural fields within 500 m were selected as controls. House dust was vacuumed in all homes from floors (VFD) and from newly placed clean doormats (DDM). Sampling was done during two periods, when pesticides are used and not-used. For determination of 46 prioritized pesticides, a multi-residue extraction method was used. Most statistical analyses are focused on the 12 and 14 pesticides that were detected in >40% of DDM and VFD samples, respectively. Mixed models were used to evaluate relationships between possible determinants and pesticides occurrence and concentrations in DDM and VFD. 17 pesticides were detected in more than 50% of the homes in both matrixes. Concentrations differed by about a factor five between use and non-use periods among homes within 250 m of fields and between these homes and controls. For 7 pesticides there was a moderate to strong correlation (Spearman rho 0.30–0.75) between concentrations in DDM and VFD. Distance to agricultural fields and air concentrations were among the most relevant predictors for occurrence and levels of a given pesticide in DDM. Concentrations in dust are overall higher during application periods and closer to fields (<250 m) than further away. The omnipresence of pesticides in dust lead to residents being exposed all year round.

Ingestion of microplastics by aquatic organisms is often harmful due to the dilution of their regular food with low-calorie microplastic particles, but can also be beneficial if nutritious biofilms are present on the microplastic surface. This begs the question: is ingestion of microplastic harmful or beneficial and can the net effect of the two mechanisms be quantified? Here, we quantified these harmful and beneficial effects on Daphnia magna, using dose-response tests with clean and biofouled microplastic respectively, and determined the trade-off between these counteracting effects. A population model was developed to calculate the isoclines for zero population growth, separating the regime where adverse food dilution dominated from that where the beneficial biofilm vector mechanism dominated. Our results show that the organisms grew better when exposed to biofouled microplastic compared to pristine microplastic. Very good model predictions (R2 = 0.868–0.991) of the effects of biofouled microplastic were obtained based on literature parameter values, with optimization required only for the two sub-model parameters driving the dose-effect relationships for pristine microplastic. These results contradict previous sudies were only pristine microplastic were used and demonstrate that the ruling paradigm of unambiguously adverse microplastic effects is not ecologically justifiable.

Ingestion of microplastics by aquatic organisms is often harmful due to the dilution of their regular food with low-calorie microplastic particles, but can also be beneficial if nutritious biofilms are present on the microplastic surface. This begs the question: is ingestion of microplastic harmful or beneficial and can the net effect of the two mechanisms be quantified? Here, we quantified these harmful and beneficial effects on Daphnia magna, using dose-response tests with clean and biofouled microplastic respectively, and determined the trade-off between these counteracting effects. A population model was developed to calculate the isoclines for zero population growth, separating the regime where adverse food dilution dominated from that where the beneficial biofilm vector mechanism dominated. Our results show that the organisms grew better when exposed to biofouled microplastic compared to pristine microplastic. Very good model predictions (R² = 0.868–0.991) of the effects of biofouled microplastic were obtained based on literature parameter values, with optimization required only for the two sub-model parameters driving the dose-effect relationships for pristine microplastic. These results contradict previous sudies were only pristine microplastic were used and demonstrate that the ruling paradigm of unambiguously adverse microplastic effects is not ecologically justifiable.

Copper (Cu), as an essential element, is added to animal feed to stimulate growth and prevent disease. The forage crop alfalfa (Medicago sativa L.) produced during Cu phytoextraction may be considered a biofortified crop to substitute the Cu feed additives for livestock production, beneficially alleviating Cu contamination in soils and reducing its input into agriculture systems. To assess this, alfalfa was grown in three similar soils with different Cu levels, i.e., 11, 439 and 779 mg kg⁻¹ for uncontaminated soil (A), moderately Cu-contaminated soil (B) and highly Cu-contaminated soil (C), respectively. EDDS (Ethylenediamine-N,N′-disuccinic acid) was applied to the soils seven days before the first cutting at four rates (0, 0.5, 2 and 5 mmol kg⁻¹) to enhance bioavailable Cu uptake. Alfalfa grew well in soils A and B but not in the highly Cu-contaminated soil. After applying EDDS, a significant biomass reduction of the first cutting shoot was only observed with 5 mmol kg⁻¹ EDDS in the highly Cu-contaminated soil, with a 45% (P < 0.05) decrease when compared to the control. Alfalfa grown in the three soils gradually wilted after the first cutting with 5 mmol kg⁻¹ EDDS, and Cu concentrations in the first cutting shoot were augmented strongly, by 250% (P < 0.05), 3500% (P < 0.05) and 6700% (P < 0.05) compared to the controls, respectively. Cu concentrations in alfalfa shoots were found to be higher in this study than in some fodder plants and further augmented in soils with higher Cu levels and with EDDS application. These findings suggest that alfalfa grown on clean soils or soils with up to 450 mg Cu kg⁻¹ (with appropriate EDDS dosages) has the potential to be considered as a partial Cu supplementation for livestock. This research laid the foundation for the integration between Cu-phytoextraction and Cu-biofortification for livestock.

International audience | Copper (Cu), as an essential element, is added to animal feed to stimulate growth and prevent disease. The forage crop alfalfa (Medicago sativa L.) produced during Cu phytoextraction may be considered a biofortified crop to substitute the Cu feed additives for livestock production, beneficially alleviating Cu contamination in soils and reducing its input into agriculture systems. To assess this, alfalfa was grown in three similar soils with different Cu levels, i.e., 11, 439 and 779 mg kg−1 for uncontaminated soil (A), moderately Cu-contaminated soil (B) and highly Cu-contaminated soil (C), respectively. EDDS (Ethylenediamine-N,N′-disuccinic acid) was applied to the soils seven days before the first cutting at four rates (0, 0.5, 2 and 5 mmol kg−1) to enhance bioavailable Cu uptake. Alfalfa grew well in soils A and B but not in the highly Cu-contaminated soil. After applying EDDS, a significant biomass reduction of the first cutting shoot was only observed with 5 mmol kg−1 EDDS in the highly Cu-contaminated soil, with a 45% (P < 0.05) decrease when compared to the control. Alfalfa grown in the three soils gradually wilted after the first cutting with 5 mmol kg−1 EDDS, and Cu concentrations in the first cutting shoot were augmented strongly, by 250% (P < 0.05), 3500% (P < 0.05) and 6700% (P < 0.05) compared to the controls, respectively. Cu concentrations in alfalfa shoots were found to be higher in this study than in some fodder plants and further augmented in soils with higher Cu levels and with EDDS application. These findings suggest that alfalfa grown on clean soils or soils with up to 450 mg Cu kg−1 (with appropriate EDDS dosages) has the potential to be considered as a partial Cu supplementation for livestock. This research laid the foundation for the integration between Cu-phytoextraction and Cu-biofortification for livestock.